BACKGROUND:Occurrence and development of diabetic complications is closely related to the severity of oxidative stress imbalance in the body. OBJECTIVE:To investigate the effect of embryonic stem cel s on oxidative stress response of rats with diabetic nephropathy. METHODS:Primarily cultured rat embryonic stem cel s were observed for cel morphology and surface antigen detection. Sprague-Dawley rats were divided into experimental group (two injections of embryonic stem cel s via the tail vein), model group (injection of the same volume of PBS), and normal control group (with no modeling, intraperitoneal injection of sodium citrate-citrate buffer). In the former two groups, the rats were intraperitoneal y injected sodium citrate-citrate buffer diluted streptozotocin to establish diabetic nephropathy models before treatment. At 5 weeks after the last injection, blood glucose level, renal function indicators (urine protein/urine creatinine, blood urea nitrogen and serum creatinine) were tested in each group;contents of malondialdehyde and protein carbonyl were detected in the kidney;the expression level of superoxide dismutase was detected by western blot assay. RESULTS AND CONCLUSION:The embryonic stem cel s were oval or round, with clear boundary and good refraction, and highly expressed Oct-4 and SSEA-1. Compared with the control group, renal biochemical indicators, malondialdehyde and protein carbonyl contents were significantly increased, while the expression level of superoxide dismutase was decreased dramatical y in the model group and experimental group (P<0.05);compared with the model group, the renal biochemical indicators, malondialdehyde and protein carbonyl contents were dropped significantly in the experimental group, but the expression of superoxide dismutase was significantly rebounded (P<0.05). Taken together, embryonic stem cel s can reverse the occurrence and development of diabetic nephropathy by inhibiting oxidative stress in progress.

Objective To determine the effects of prostaglandin E_2 alone and the combination with omeprazole for poisoning related acute mucosal lesion. Methods Forty-two healthy SD rats were randomly divided into 7 groups: control group (control), methamidophos poisoning model group (intox), 10 mg/kg omeprazole group (ome1), 50 mg/kg omeprazole group (ome2), 10 μg/kg prostaglandin E_2 group (PG1), 50 μg/kg prostaglandin E_2 group (PG2), 10 μg/kg prostaglandin E_2 combined with 10 mg/ kg omeprazole group (union). The gastric ulcer index was counted and the content of serum cholinesterase, endothelin, and tumor necrosis factor were measured, and dependability was analyzed. Results There was no obvious difference between the prostaglandin group and the poisoning model group in gastric ulcer index, endothelin, and tumor necrosis factor. The omeprazole alone and the combination treatment could improve the acute gastric mucosa lesion induced by methamidophos poisoning. Conclusion Prostaglandin E_2 alone can not obviously improve the acute gastric mucosa lesion by methamidophos poisoning. The combination with omeprazole may be a promising treatment.

Objective To observe the changes of Endothelin,Tumor Necrosis Factor in different stress; and to explore the corelation between the changes of Endothelin,Tumor Necrosis Factor and the occurrence of stress-related gastric mucosal disease.Methods The models of the acute methamidophos poison and the acute myocardial infarction in rats were made.The gastric ulcer index was counted and the content of blood plasma Endothelin and blood serum Tumor Necrosis Factor were checked.Results Models of the acute methamidophos poison and the acute myocardial infarction in rats were set up successfully. The gastric ulcer index of rats in the stress groups increased.The blood plasma Endothelin and blood serum Tumor Necrosis Factor in the stress group were significantly higher than those in the control group.Conclusion The gastric mucosal disease under different stress was observed with the gastric mucosal ulcer index.The blood plasm Endothelin and blood serum Tumor Necrosis Factor under different stress were increased,suggesting they play an important role in this phenomenen.

0. 05), the mean effective discharge time was 3. 4+ 3. 2 (P0. 05), the mean effective discharge time was 2. 6+1. 7 (P

BACKGROUND: The transplantation of allogeneic cartilage has local immunological rejection, and it is necessary to further reduce the rejection to promote its application in clinic, thus it is significant to perform a series of experiments to induce local immune privilege.OBJECTIVE: To observe the in vivo growth of tissue engineered allogeneic cartilage reconstructed by chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL.DESIGN: A randomized controlled observation.SETTING: Shanghai Jiao Tong University.MATERIALS: Thirty-six allogeneic New Zealand rabbits as recipients and 45 1-week-old chinchillas as donors, either sex,were purchased by the experimental animal center of Chinese Academy of Sciences. Amphotropic recombinant retrovirus coated cell line PT67 was purchased from Clontech Company; Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), G418 and Polybrene were bought from GIBCO BRL.METHODS: The experiment was carried out in original Shanghai Second Medical University from January 2000 to July 2005. The New Zealand rabbits were randomly divided into three groups: FasL-transfected group (n =12), untransfected group (n =12) and blank control group (n =12). The rabbit allogeneic cartilages were constructed by the compound of pLNCX2-FasL transfected chondrocytes and tissue engineered material of pluronic F-127. ① Gross observation and mass changes of the grafts: Corresponding materials were infused subcutaneously, the grafts were removed at 1, 2 and 3 months after transplantation for gross observation and the mass changes. ② Staining observation: The grafts were removed at 1, 2 and 3 months after transplantation, then prepared into sections, and observed by hematoxylin and eosin (HE), Safranin'O and Masson's trichrome stainings. ③ Antibody detection: Blood samples (1 mL) were collected at 1 and 2 months after transplantation, the chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, and separated by electrophoresis in agarose medium, then acted with serum of recipient to observe whether corresponding antibody generated. ④ Complement dependent cytotoxicity (CDC) test: The chondrocytes of chinchillas were prepared into cell suspension (2×109/L), and then seeded into 96-well plate, attached grew for 24 hours, then recipient serum was added for the CDC test, and the percentage of apoptotic cells was counted under microscope.MAIN OUTCOME MEASURES: ① Gross observation and mass changes of the grafts;② Histological changes; ③ Results of the antibody detection; ④ Percentage of apoptotic cells.RESULTS: All the 81 rabbits were involved in the analysis of results. ① Gross observation and mass changes of the grafts: Two weeks after inoculation, there were obvious nod formations at the inoculated sites, but no nod formed in the blank control group. The new cartilage tissues became smaller gradually and completely disappeared at 4 months in the untransfected group, whereas those in the FasL-transfected group became smaller, but still existed after 7 months. The masses of grafts in the FasL-transfected group were higher than those in the untransfected group (P ＜ 0.05). ②Histological observation: Plenty of lymphocytic infiltrations around cartilage tissue could be observed in the untransfected group, and obviously decreased in the FasL-transfected group. No lymphocyte was observed inside the chondrocytes.Masson's trichrome staining was performed, and it was observed under light microscope that the small white parts in the middle were immature chondrocytes, and there were green collagen around most of the mature chondrocytes. Safranin O staining showed strong positive reaction, suggested that there were rich glycosaminoglycan in matrix. ③ Antibody detection: The chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, then acted with serum of recipient, and the results showed that no corresponding antibody generated. ④ Percentage of apoptotic cells: The percentages of serum CDC apoptotic cells in the FasL can ransfected group, untransfected group and blank control group were 5%, 6% and 1%, which were all negative.CONCLUSION: Rabbit allogeneic chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL can reconstruct tissue engineered cartilage, and can postpone the degeneration by 3 months.

BACKGROUND: At present,enhanced green fluorescent protein(EGFP)is proved to be the best labeled molecule,with unique advantages,such as high fluorescence specificity,easy to be detected,and so on.Recombined retroviral vector EGFP-pLNCX2,which can stably express EGFP,can be construct using gene-engineering technique.Transfecting allogenic chondrocytes is indeed very useful to investigate the target gene expression and process of constructing tissue engineered cartilage in vivo.OBJECTIVE:To construct recombined retroviral vector EGFP-pLNCX2 which can stably express EGFP,and investigate optimal conditions for retrovirus transfection of chondrocytes.DESIGN:Randomized controlled observation.SETTING:Shanghai Jiao Tong University.MATERIALS:Thirty New Zealand rabbits of either gender,1 week of age,were purchased from Experimental Animal Center of Chinese Academy of Sciences.Amphotropic retrovirus package cell line PT67 pLNCX2 and pEGEP-C1 were purchased from Clontech Company;NIH 3T3 cell line was purchased from ATCC Company;DH5a Bacterium coli was preserved by the laboratory of Shanghai Jiao Tong University;Retroviral vector pLNCX2 was purchased from Clontech Company;pEGFP-C1 plasmid with EGFP were donated by professor Cong Xiao-qian from Chinese Academy of Sciences.METHODS:This experiment was carried out in the Shanghai Jiao Tong University in August 2005.Chondrocytes of New Zealand rabbits were isolated and cultured.The recombinant retroviral vector EGFP-pLNCX2,which can stably express EGFP,was constructed to transfect cultured chondrocytes from New Zealand rabbits by using gene engineering technique.Transfection results were observed under fluorescence microscope.Altogether 6×105 chondrocytes incubated in 10 cm-diameter flat plate were used to transfect retrovirus-EGFP immediately and at 12,24 and 48 hours after inoculation.One week later,EGFP expression efficiency was measured with flow cytometer,and best occasion for retrovirus transfecting primary chondrocytes.Enzyme-digested chondrocytes were inoculated for 24 hours to transfect retrovirus.After 250 mg/L G418 was added,chondrocytes were screened on the 2nd,3rd,4th,5th and 6th days separately.After phosphate buffer solution(PBS)washings,the transfection efficiency of chondrocytes was detected by flow cytometer,and the best occasion for G418 screening was observed.MAIN OUTCOME MEASURES:①EGFP-pLNCX2 transfection efficiency.②The best occasions for retrovirus transfecting primary chondrocytes and G418 screening.RESULTS: ①Retroviral vector EGFP-pLNCX2 transfected primary chondrocytes of rabbits,and high expression of transfected chondrocytes could be obtained through preliminary screening of G418.After being screened and expressing EGFP,chondrocytes kept normal morphology,with pseudopod adhering to the wall and matrix secreting vigorously.②The best occasion for retrovirus transfecting primary chondrocytes was at 24 hours after cell inoculation.The transfection efficiency determined with flow cytometer was 19.14% on the 5th day.The best occasion for G418 screening was on the 5th day after culture.Transfection efficiency of G418 screening was 55.75% on the 7th day.CONCLUSION:Recombinant retroviral vector EGFP-pLNCX2 can effectively transfect chondrocytes.The best occasion for retrovirus transfecting primary chondrocytes is at 24 hours after inoculation, and the best occasion for G418 screening is on the 5th day after culture.

Objective To investigate the current situation of healthcare-associated infection(HAI)management in secondary and above medical institutions in Shaanxi Province,analyze development trend,and put forward sugges-tions for improvement.Methods In May-June,2016,170 secondary and above hospitals in 10 cities were selected for surveying through stratified random sampling method.Survey content included basic situation of hospitals,HAI management,HAI monitoring,and so on.Results Available questionnaires were obtained from 165 hospitals (43 tertiary hospitals,and 122 secondary hospitals).Of 165 hospitals,more than 90% have established HAI manage-ment organizations and regulations,but hospital risk management should be paid more attention,only 63.03% of hospitals perfected the risk management system and 66.06% conducted risk assessment.99.09% of hospitals im-plement training on HAI to all staff regularly and 88.41% conducted effective feedback.In the aspect of staff alloca-tion,88.48% of the hospitals assigned enough professionals for HAI management,but only 34.55% have specific training programme for these personnel.Only 33.94% of hospitals have special funds for HAI control;in the aspect of monitoring on HAI,21.21% of hospital installed and used HAI monitoring software;In the aspect of implemen-tation of monitoring programme,about 90% of hospitals developed monitoring on HAI cases and environmental hy-giene,but only 34.55% and 23.64% of hospitals conducted targeted monitoring on intensive care unit and neonatal intensive care unit respectively.Conclusion Organizational structure of HAI management in Shaanxi Province is perfect,relevant rules and regulations are basically established,basic monitoring projects are universal,but the awareness of risk management needs to be strengthened,professional allocation and professional quality develop-ment are both imbalance,informational monitoring is inadequate.

OBJECTIVE:To evaluate the effect of psychological intervention based on pharmaceutical care on the communi-ty patients with essential hypertension. METHODS:172 patients with essential hypertension in a community health service certer in Nanjing Gulou area in 2013 were divided into control group(n=87)and intervention group(n=85)by simple random meth-od. Control group received conventional antihypertensive drugs and management,intervention group additionally received pharma-ceutical care and psychological intervention,it lasted 6 months. Blood pressure controlling,drug compliance,mastery of drug knowledge and health status in 2 groups before and after treatment were compared. RESULTS:Before treatment,there were no significant differences in the systolic blood pressure(SBP)and diastolic blood pressure(DBP)between 2 groups(P>0.05);af-ter treatment, SBP and DBP in 2 groups were lower than before,SBP and DBP in intervention group were lower than control group,the decrease degree of SBP and DBP was bigger than control group,the differences were statistically significant (P<0.01). 1 month after the beginning of intervention,there was no significant difference in the proportion of drug compliance be-tween 2 groups (P>0.05);1 month after the end of the intervention,the proportion of drug compliance in intervention group was higher than 1 month after the beginning of intervention and control group,the differences were statistically significant(P<0.05). Before treatment,there was no significant difference in mastery of drug knowledge between 2 groups (P>0.05);after treatment,the proportion of patients showed good and excellent mastery of drug knowledge was higher than before and control group,the differences were statistically significant (P<0.05). Before treatment,there were no significant differences in scores of physical,mental,social health subscale and SRHMS between 2 groups(P>0.05);after treatment,scores of physical health subscale and SRHMS in control group,each subscale and to-tal scale scores in intervention group were higher than be-fore, and scores of mental, social health subscale and SRHMS in intervention group were higher than control group, the differences were statistically significant (P<0.05). CONCLUSIONS: The psychological intervention based on pharmaceutical care can effectively promote the con-trol of blood pressure,improve drug compliance,mastery of drug knowledge,health status and mental situation,which is worth of popularizing and applying in community health services.

Objective To investigate the effect of estrogen on the pain score,c-Fos and substance P expressions in the dorsal horn of the spinal cord in the mice following formalin stimulation.Methods Fifteen C57/BL6 mice were randomized to 3 groups: control group(intact mice without estrogen treatment),OVX+V group(ovariectomized mice given vehicle) and OVX+E group(ovariectomized mice with subcutaneous injection of 2 ?g/d 17?-estradiol for 10 d).Pain score was used to assay the role of estrogen in affecting pain threshold in the mice following formalin injected into the right hind paw,and expressions of c-Fos and substance P in the dorsal horn of spinal cord(L3 to L5) in 2 h after injection of formalin was tested with immunohistochemisty to evaluate neuron activity and pain afferent fibers.Results Pain score was increased in ovariectomized mice following formalin stimulation,which was inhibited by estrogen especially in the early stage of secondary phase.The number of c-Fos-like immunoreactivity neuron(FLIN,P

Objective To explore the effect of modified Zexie Decoction on renal aquaporin-2 (AQP2) expression in high-salt hypertensive rat. Methods Hypertensive rats model was established by feeding rat with 8%high salt. Rats (n=50) were divided into model group, modern medicine group, traditional Chinese medicine groups of high, medium, low dose, with 10 rats in each group. The other 10 rats were fed with ordinary diet as normal group. Rats in traditional Chinese medi?cine of high, medium, low groups were given Zexie Decoction suspension of 16.2, 10.8 and 5.4 g/(kg·d) respectively;Rats in modern medicine group was given Valsartan hydrochlorothiazide 16.65 mg/(kg·d);the model group and normal group was ad?ministered with equal volume of distilled water. Animals were feed with medications at 1 mL/100 g by gavage for 4 weeks. On the 1st , 4th , 7th , 14th and, 28th day of administration, we measured SBP and collected 24 h urine. We employed immunohis?tochemistry to detect renal AQP-2 protein expression level and RT-PCR to detect renal AQP-2 mRNA transcription level. Results The rank of SBP from high to low is:model group>traditional Chinese medicine medium and low dose groups>traditional Chinese medicine high dose group and western medicine group>normal group. The rank of urine volume from high to low is:Western medicine group and traditional Chinese medicine high dose group>traditional Chinese medicine me?dium and low dose group > normal group, the difference was not statistically significant between traditional Chinese medi?cine medium and low dose group , or between western medicine group and traditional Chinese medicine high dose group. The renal AQP-2 in epithelial cells along the collecting duct wall of rats in model group show brown particles which are darker and wider distributed than those in normal group and traditional Chinese medicine of high, medium, low dose groups. RT-PCR results show that AQP-2 mRNA expression is highest in rats of model group and lowest in rats of traditional Chi?nese medicine high dose group (P<0.05). No statistical significance of AQP mRNA level was found between traditional Chi?nese medicine low group and model group (P < 0.05). Conclusion Modified Zexie Decoction can lower blood pres?sure by inhibiting the expression of AQP-2.